基于COMSTAT软件对微生物被膜进行定量分析的方法与意义

目的:微生物被膜是一种具有协调性、功能性和高度结构性的膜状复合物,它可以为微生物提供良好的生存环境,免受外界因素的干扰。研究发现,具有产生微生物被膜能力的细菌治病性明显增强,而现今缺乏一种分析微生物被膜的有效手段。本文探讨利用COMSTAT软件对微生物被膜进行定量分析的方法和意义,为对微生物被膜定性定量分析提供支持手段,从而为研究微生物被膜致病性提供方法理论基础。方法:以葡萄球菌为研究模型,利用激光扫描共聚焦显微镜成像技术,结合COMSTAT微生物被膜分析软件对微生物被膜的单位面积生物量、基质覆盖率、平均厚度、粗糙系数等方面进行定量分析,研究了该葡萄球菌的生物被膜生长变化过程,并考察了抗生素对其生物被膜的抑制作用。结果:在葡萄球菌生物被膜生长过程中,生物量、平均厚度以及平均扩散距离等结构指标数值都有明显增加,而粗糙度和表面积与生物量比值呈现降低趋势,表明了微生物被膜由发生向成熟的转化过程。与此同时,经10μg/mL和100μg/mL的卡那霉素处理得到的葡萄球菌微生物被膜生长受到明显抑制,且随着卡那霉素的浓度增加,抑制效果随之增加。结论:本文运用COMSTAT软件的分析方法首次从生物量、平均厚度等结构指标数值的角度描述了葡萄球菌生物被膜,从而有效评价微生物被膜发生、发展、成熟以及崩解的生长过程。该技术在研究微生物被膜形成的理论机制方面存在潜在价值,可以为研究微生物被膜治病性提供理论基础,具有理论指导意义。

Construction and Significance of Quantitative Analysis of Microbial BiofilmBased on COMSTAT Program

Objective： Microbial biofilms are coordinated, functional and highly structured community. Under the protection of biofilms, bacteria can be free from the interference of external factors. Studies found that biofilms should be responsible for some chronic and incurable diseases caused by pathogenic bacteria. Therefore, a pressing matter is searching for effective methods for the analysis of bacterial biofilm. We investigate the construction and significance of quantitative analysis of microbial biofilm based on COMSTAT pro gram and aim to provide the method for qualitative and quantitative analysis, as well as theoretical basis of biofilm of pathogenic microorganisms. Methods： According to confocal microscopy imaging combined with COMSTAT analysis, Staphylococcus sp. was selected as the object of study. Features of Bio-volume, average thickness, average diffusion distance and etc. were determined to charac terize the growth process of biofilm and inhibition of kanamycin. Results： During the biofilm process of Staphylococcus sp. feature index ofbio-volume, average thickness and average diffusion distance are significantly increased, while the roughness coefficient and surface to volume ratio showed a decreasing trend, indicated the maturation process of biofilm. Also, biofilm structure of Staphylococcus sp. was inhibited by Kanamyein in both 10 μg/ml and 100 μg/ml treatment. Conclusion： The structural index of Staphylococcus sp. is well characterized, and the occurrence, development, maturation and dissolution process of biofilm are well evaluated based on quantitative analysis of COMSTAT program. There existed potential value on the study of theory mechanism of microbial biofilm formation, and could provide theoretical basis for the research on microbial biofilm treatment.